环巴胺进入Ⅰ期临床试验

环巴胺(Cyclopamine)是一种异甾体类生物碱,主要存在百合科植物中北美山藜芦(Veratrum californicum)、印第安鹿食草(Cornlily)及毛叶藜芦(Veratrum grandiflorum)、伊贝(Fritillaria pallidiflora Schrenk)四种植物中。二十世纪中期研究发现环巴胺具有致畸作用,但九十年代以后的研究表明,环巴胺是一种hedgehog信号通路抑制剂,由于hedgehog信号通路的突变与多种肿瘤的发病有关联,因此环巴胺作为一种潜在的抗肿瘤新药在世界范围内掀起了研究热潮。环巴胺对髓质母细胞瘤、基底细胞癌、小细胞肺癌有较好抑制作用,对老年性前列腺癌、泌尿系统…     

环巴胺固体脂质纳米粒的研究

目的 制备环巴胺固体脂质纳米粒(Cyc-SLNs),并考察其理化性质和对Hedgehog信号通路的抑制作用.方法 采用薄膜超声法制备Cyc-SLNs,并考察其对B16F10黑色素瘤细胞增殖、细胞周期、凋亡和Hedgehog信号通路靶基因Gli1表达的影响.结果 制得的Cyc-SLNs呈平整的球形,平均粒径、多分散指数(PDI)、Zeta电位和平均包封率分别为126.5 ±2.1 nm、0.198±0.007、23.5±0.7 mV、91.59％±0.37％;Cyc-SLNs对B16F10细胞增殖分裂的抑制、凋亡的促进和Gli1表达的下调均强于环巴胺.结论 成功制备了抗肿瘤细胞效果强于环巴胺的固体脂质纳米粒Cyc-SLNs.     

黑紫藜芦化学成分研究

目的 研究黑紫藜芦(Veratrum japonicum Loes.f.)的化学成分.方法 利用酸碱处理及硅胶柱色谱进行系统分离,通过理化性质和波谱数据鉴定结构;利用黄鸣龙反应还原主要成分介藜芦碱.结果 分离并鉴定了其中5个化合物,分别是正二十八烷醇(1)、β-谷甾醇(2)、表红介藜芦碱(3)、胡萝卜苷(4)、介藜芦碱(5);还原介藜芦碱得到抗肿瘤活性物质环巴胺.结论 化合物1～4均为首次从该植物中分离得到;首次利用黄呜龙反应还原介藜芦碱,并得到一个副产物3-表环巴胺.     

铁离子对环吡酮胺抑制犬小孢子菌的影响

目的:探索抗真菌药物环吡酮胺可能的作用机制。方法:利用微量液基稀释法和纸片扩散法,测定抗真菌药物环吡酮胺抑制犬小孢子菌时铁离子的作用。结果:所有受试菌株均表现为对环吡酮胺耐药。结论:铁离子参与了抗真菌药物环吡酮胺对犬小孢子菌的抑菌作用。     

注射用环磷腺苷葡胺细菌内毒素检查

目的对注射用环磷腺苷葡胺进行凝胶法干扰试验,建立注射用环磷腺苷葡胺的细菌内毒素检查试验方法。方法按照《中国药典)2010年版(二部)附录ⅪE进行实验和结果判断。结果注射用环磷腺苷葡胺稀释至浓度0.15mg.mL-1时对细菌内毒素检查法无干扰作用。结论使用细菌内毒素检查法检查注射用环磷腺苷葡胺中的细菌内毒素是可行的,可用细菌内毒素检查法代替家兔热原检查法。     

老年高血压治疗新方法

其实,吲达帕胺缓释剂(纳催利缓释剂)和美利巴、寿比山属一类药物,只是它是缓释剂型,具有更长效的降压作用,而美利巴和寿比山属于普通剂型.吲达帕胺缓释剂属于长效低毒口服利尿药,通过干扰肾小管对电解质的重吸收,使钠、钾、氯和水排泄增加,但其化学结构与常用的利尿药噻嗪类不同.     

对环磷腺苷葡胺注射剂含量测定方法的探讨

目的:环磷腺苷葡胺注射剂是由环磷腺苷与葡甲胺按摩尔比1:1加适量辅料制成,现行标准中均规定测定环磷腺苷葡胺的含量,认为环磷腺苷与葡甲胺结合为单一化合物。对现行标准的含量测定方法进行考察,研究含量测定计算方法的合理性。方法:对现行标准中的液相色谱法进行验证,考察环磷腺苷和葡甲胺在现行检验标准的色谱条件下的色谱行为。结果:现行标准采用的10种流动相体系中,环磷腺苷和葡甲胺为两个单独的色谱峰,并无单独的环磷腺苷葡胺峰。目前所有的含量测定方法中以环磷腺苷为对照品,计算环磷腺苷葡胺含量都是不合理的,环磷腺苷和葡甲胺应分别计算。     

环磷腺苷葡胺与磷酸肌酸钠治疗新生儿窒息后心肌损害的临床观察

目的观察环磷腺苷葡胺与磷酸肌酸钠治疗新生儿窒息后心肌损害的治疗效果。方法将58例新生儿窒息后心肌损害的患儿随机分成磷酸肌酸钠(34例)与环磷腺苷葡胺组(24例),两组病例均按照新生儿窒息作综合处理,磷酸肌酸钠组加用磷酸肌酸钠静脉滴注,剂量为0.5g/次,1次/d,环磷腺苷葡胺组加用环磷腺苷葡胺静脉滴注,剂量为3~5mg/(kg次),1次/d,7d为1疗程。观察两组患儿治疗后症状、体征、肌酸激酶同工酶CK-MB和肌钙蛋白I(cTnI)的改变。结果环磷腺苷葡胺组与磷酸肌酸钠组在有效率、改善CK-MB和cTnI方面临床结果相近。结论磷酸肌酸钠和环磷腺苷葡胺作用于新生儿窒息后心肌损伤的不同环节,临床观察对改善心脏泵血功能、保护心肌细胞疗效相似,其预后相近,但环磷腺苷葡胺价格不及磷酸肌酸钠的1/5,能显著减轻患者负担,有临床推广价值。     

新霉胺类似物的合成与RNA结合和生物活性评价

为了提高新霉胺对16S rRNA的亲和力，合成了Ⅱ环5位修饰的新霉胺类似物。以新霉素B为原料，经水解，保护，亲核取代，脱保护，叠氮还原多步反应得到氨基或氨基链修饰的新霉胺类似物。用表面等离子共振法测定了所合成的化合物与大肠杆菌（E.coli．）核糖体A位点rRNA（16S RNA）的相互作用。合成了6个Ⅱ环5-位修饰的新霉胺类似物，发现Ⅱ环5位氨基链修饰可以增强化合物对16S RNA的亲和力，其中一些化合物在10^-3M有体外细菌抑制活性。在新霉胺的Ⅱ环5位引入氨基或脂肪胺可以增加与16S RNA的亲和力。Ⅱ环5位上羟基的构型改变对于药物／16S RNA复合物稳定性的影响较低。     

环磷腺苷葡胺联合依那普利对慢性心力衰竭患者BNP的影响分析

目的探讨环磷腺苷葡胺联合依那普利对慢性心力衰竭患者血浆脑钠肽(BNP)的影响。方法将120例慢性心力衰竭患者随机分为两组,对照组各60例使用环磷腺苷葡胺治疗,研究组各60例使用环磷腺苷葡胺联合依那普利治疗,比较两组治疗前后血浆BNP水平及临床疗效。结果治疗后,研究组血浆BNP水平明显低于对照组(P <0.05)。研究组有效率为95.0%,明显高于对照组的83.3%(P <0.05)。结论慢性心力衰竭患者采用环磷腺苷葡胺联合依那普利治疗可有效降低血浆BNP水平,提升疗效。     

Hedgehog antagonist cyclopamine isomerizes to less potent forms when acidified

The effect of acid treatment of cyclopamine, a natural antagonist of the hedgehog (Hh) signaling pathway and a potential anti-cancer drug, has been studied. Previous reports have shown that under acidic conditions, as in the stomach, cyclopamine is less effective. Also, it has been stated that cyclopamine converts to veratramine, which has side effects such as hemolysis. In this study, we examined in detail the influence of acidification on structure and activity of cyclopamine. We found that of acidified cyclopamine converts to two previously unreported isomers, which we have called cyclopamine (S) and cyclopamine (X). These have likely gone undetected because cyclopamine is often analyzed with fast and hence lower resolving chromatographic methods. Compared to natural cyclopamine, these cyclopamine isomers have a significantly reduced effect on the ciliary transport of the Hh receptor smoothened, and reduced inhibition on the Hedgehog signaling pathway. The side effects of these isomers are unknown. Our findings can partly explain a reduced efficiency of cyclopamine in a gastric environment, and may help with the rational design of more pH independent cyclopamine analogues.     

Agents targeting the Hedgehog pathway for pancreatic cancer treatment

Recent evidence has demonstrated that aberrant reactivation of the Hedgehog signaling pathway contributes to tumor initiation and progression in various human malignancies, including pancreatic cancer; therefore, the Hedgehog pathway has emerged as a promising novel therapeutic target. Initial translational studies conducted using cyclopamine, a small-molecule inhibitor of the Smoothened (SMO) component of the Hedgehog pathway, demonstrated that pharmacological blockade of aberrant Hedgehog signaling has the potential to inhibit tumor initiation, progression and metastatic spread. This concept has been corroborated using different compounds in various preclinical models of pancreatic cancer and other malignancies; several of these studies suggest possible therapeutic synergisms of Hedgehog inhibitors with established antineoplastic agents. This review provides a concise overview of translational studies assessing the use of Hedgehog inhibitors as novel therapeutic strategy for cancer, particularly pancreatic cancer.     

Semisynthetic cyclopamine analogues as potent and orally bioavailable hedgehog pathway antagonists

Herein is reported the synthesis of a novel class of hedgehog antagonists derived from cyclopamine. The acid sensitive D-ring of cyclopamine was homologated utilizing a sequence of chemoselective cyclopropanation and stereoselective acid-catalyzed rearrangement. Further modification of the A/B-ring homoallylic alcohol to the conjugated ketone led to the discovery of new cyclopamine analogues with improved pharmaceutical properties and in vitro potency (EC 50) ranging from 10 to 1000 nM.     

特异性阻断Shh信号通路对胃癌细胞系SGC-7901增殖和凋亡的影响

目的:研究Sonic Hedgehog(Shh)信号通路对胃癌细胞系SGC-7901增殖和凋亡的影响。方法:用四唑蓝(MTT)比色试验检测Shh信号通路特异性抑制剂cyclopamine对胃癌细胞系SGC-7901增殖的抑制作用;流式细胞术检测细胞增殖指数(PI)和凋亡指数(AI)。结果:Cyclopamine对SGC-7901细胞株增殖的抑制作用呈剂量和时间依赖性;Cyclopamine作用后使胃癌细胞周期阻滞在G0/G1期,细胞凋亡增加。结论:Shh信号分子对胃癌细胞的增殖起维持作用;通过特异性阻断该信号通路,可以抑制胃癌细胞增殖,并促进细胞凋亡。     

Cyclopamine‐induced synophthalmia in sheep: defining a critical window and toxicokinetic evaluation

Cyclopamine, a steroidal alkaloid, from the plant Veratrum californicum is teratogenic, causing a range of different birth defects. The critical window for cyclopamine‐induced synophthalmia formation has been reported to be gestational day (GD) 14. The objectives of this study were to better describe cyclopamine‐induced craniofacial deformities, to better define the window of susceptibility to synophthalmia formation, and to characterize cyclopamine toxicokinetics in sheep. Ewes were dosed i.v. with purified cyclopamine for toxicokinetic analysis. Another four groups of ewes were dosed orally twice daily with 0.88 g/kg of V. californicum on GD 13, 14 or 15 or consecutively on GD days 13–15. Pregnancy and pre‐partum fetal malformations were determined by ultrasound imaging on GD 60. At parturition lambs were assessed for gross malformations. The elimination half‐life of cyclopamine in ewes was determined to be 1.1 ± 0.1 h. The rapid clearance of cyclopamine indicates that ingestion of V. californicum must occur during a very narrow window for synophthalmia formation to occur. Ewes dosed with V. californicum on GD 13 or 14 had lambs with various craniofacial malformations including cyclopia, maxillary dysplasia and mandibular micrognathia. Ewes dosed on GD 15 delivered normal lambs. Ewes dosed consecutively on GD 13–15 were not pregnant at GD 60 and Veratrum‐induced embryonic death was assumed to be the cause. Interestingly, lambs with cyclopia were smaller, under‐developed and appeared premature even though their twin appeared fully developed. Initial evaluations suggest this was due to placental dysplasia. Published in 2009 by John Wiley & Sons, Ltd.     

Identification and characterization of several dietary alkaloids as weak inhibitors of hedgehog signaling.

The Hedgehog (Hh) signaling pathway plays an integral role in the patterning and development of diverse structures in the vertebrate embryo. Aberrations in Hh signaling are associated with a range of developmental defects including failure of interhemispheric division of the embryonic forebrain as well as midline facial dysmorphia including cleft lip/palate and cyclopia, collectively termed holoprosencephaly (HPE). Postnatally, Hh signaling has been postulated to play a pivotal role in healing and repair processes and inappropriate Hh pathway activation has been implicated in several types of cancers. The Veratrum alkaloid cyclopamine is a potent inhibitor of Hh signaling and causes HPE-like defects in diverse species including sheep, hamster, mouse, and zebra fish. Using murine cell-based assays, we have determined that a number of dietary alkaloids similar in structure to cyclopamine also inhibit Hh signaling but with significantly lower potency. We found that these dietary compounds act additively through a mechanism similar to cyclopamine, downstream of Ptc1 and upstream of Gli1. Using an embryonic zebra fish developmental assay, we found that while cyclopamine exposure caused HPE-like defects, exposure to one of these dietary compounds, solanidine, did not.     

Hedgehog antagonists cyclopamine and dihydroveratramine can be mistaken for each other in Veratrum album

A toxic plant, Veratrum album (ssp. viriscens), was found to have an inhibitory effect on Hedgehog (Hh), a developmental signaling pathway that has been shown to be active during development, in adult stem cells and in numerous human tumors. Based on earlier studies it was believed that the known Hh inhibitor cyclopamine was present in V. album (ssp. viriscens). Here we show that instead of cyclopamine, dihydroveratramine (DHV) was found in V. album (ssp. viriscens). These compounds are easily mistaken for each other, as both substances share the same molecular weight, and the same main MS/MS fragments. DHV was found to be a less potent Hh inhibitor compared to cyclopamine. This is the first reported occurrence of DVH in nature.     

Blockade of sonic hedgehog signal pathway enhances antiproliferative effect of EGFR inhibitor in pancreatic cancer cells

Aim： To investigate the expression of sonic hedgehog （SHH） and epidermal growth factor receptor （EGFR） signal molecules in pancreatic cancer cells, and to assess the inhibitory effects through the blockade of the SHH and EGFR signaling pathways by cyclopamine and Iressa, respectively. Methods： The expression of SHH and EGFR in pancreatic cancer cell lines （PANC- 1, SUIT-2, and ASPC- 1） was detected by RT-PCR and Western blot analysis. After treatment with different concentrations of cyclopamine, alone or in combination with Iressa, the antiproliferative effect on pancreatic cancer cells was analyzed by methyl thiazolyl tetrazolium assays. A flow cytometry analysis was used to detect the cellular cycle distribution and apoptosis of pancreatic cancer cells. Results： All of the 3 pancreatic cancer cell lines expressed SHH, Smoothened （SMO）, and EGFR. Cyclopamine could downregulate the expression of EGFR in all cell lines. Cyclopamine or Iressa could induce a growth inhibitory effect in a dose-dependent manner. Moreover, the combined use of 2.5 μmol/L cyclopamine and 1 μmol/L Iressa induced an enhanced inhibitory effect and a greater apoptosis rate than any agent alone. The percentage of the cell population of the G0/G1 and sub-G1 phases was significantly increased along with the increasing dose of cyclopamine and/or Iressa. Conclusion： The blockade of the sonic hedgehog signal pathway enhances the antiproliferative effect of the EGFR inhibitor through the downregulation of its expression in pancreatic cancer cells. The simultaneous blockade of SHH and EGFR signaling represents possible targets of new treatment strategies for pancreatic carcinoma.     

四川地区抗肿瘤药物风险分级现状及需求研究

目的:了解四川地区抗肿瘤药物风险分级现状及需求,以构建抗肿瘤药物风险提示系统,提高抗肿瘤药物临床应用的安全性和有效性。方法:采用调查问卷了解四川地区抗肿瘤药物风险分级现状及需求。结果:受访医务人员所在医院的抗肿瘤药物使用现状：63.29%存在抗肿瘤药物的超说明书用药,77.22%有抗肿瘤药物处方前置审核系统,82.28%为一线医师开具抗肿瘤药物处方需要上级医师审核,91.14%为配置静脉用抗肿瘤药物使用生物安全柜,86.08%为医务人员在配置抗肿瘤药物时穿戴防护用具,78.48%为抗肿瘤药物高危标识嵌入电子信息系统,81.01%有抗肿瘤药物的分级管理制度,81.01%有抗肿瘤药物分级管理目录;受访医务人员对抗肿瘤药物的风险因素及风险防范的认知程度一般,而医师、护士和药师对抗肿瘤药物风险点的风险程度认知有所不同;受访医务人员希望构建抗肿瘤药物风险提示系统并建议了相关影响因素的重要性。结论:四川地区抗肿瘤药物风险分级现状较好,受访医务人员对肿瘤药物的风险因素及风险防范的认知程度侧重于其工作岗位,强烈要求构建抗肿瘤药物风险提示系统。